Roles of free fatty acid macrophage mediated killing of mycobacteria

游离脂肪酸巨噬细胞介导的分枝杆菌杀伤作用

• 批准号: 13670272
• 负责人: TOMIOKA Haruaki
• 金额: $ 2.43万
• 依托单位: Shimane medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670272/
• 关键词: mycobacterium; macrophage; effector molecule; free fatty acid; phospholipase A_2; phagosome; Mycobacterium tuberculosis; M.avium complex; ホスホリパーゼA_2; ファゴゾーム

We previously reported that reactive nitrogen intermediates and free fatty acid (FFA) play important roles in the expression of the activity of mouse peritoneal macrophages (Mφs) against Mycobacterium tuberculosis (MTB) and M. avium complex (MAC). In the present study, we examined the roles of phospholipase A_2 (PLA_2) which specifically releases arachidnic acid (AA) from membrane phospholipid in Mφantimycobacterial activity, and obtained the following results. (1) The intracellular growth of MTB residing in IFN-γ-activated Mφs was accelerated by the quinacrine (PLA_2 inhibitor) and a-TFMK (cPLA_2 inhibitor) (2) The radioactivity of MTB organisms, which were recovered from ^3H-AA loaded Mφs, was progressively increased during Mφ cultivation after MTB infection. Such a phenomenon was blocked when Mφs were treated quinacrine or a-TFMK, indicating that AA translocated to MTB within phagosome in a cPLA_2-dependent fashion. (3) The expression of cPLA_2 and iNO5 mRNAs was increased in IFN-γ- activated Mφ, this is not the case for sPLA_2 mRNA. (4) Fluorescence microscopy on infected Mφs stained with anti cPLA_2 antibody and pyrene labelled cPLA_2 substrates demonstrated that cPLA_2 translocated to mycobacterial organisms within the phagosomes of the Mφs. (5) In the case of mouse peritoneal Mφs, MAP kinase-dependent cPLA_2 phospholylation at the serine residues was observed in the early phase of Mφ cultivation after MTB infection, gradually increased at least for 48 h. On the other hand, constitutive phospholylation of cPLA_2 was observed in RAW Mφs regard less of MTB infection. These findings indicate that cPLA_2 plays important roles in the expression of antimicrobial activity of Mφs against MTB and MAC.

我们曾报道活性氮中间体和游离脂肪酸（FFA）在小鼠腹腔巨噬细胞（Mφs）抗结核分枝杆菌（MTB）和M.鸟复合体（MAC）。本实验研究了磷脂酶A_2（PLA_2）在Mφ抗分枝杆菌活性中的作用。(1)IFN-γ激活的Mφ中MTB的细胞内生长可被奎纳克林（PLA_2抑制剂）和α-TFMK（cPLA_2抑制剂）加速。（2）从负载^3 H-AA的Mφ中回收的MTB生物体的放射性在感染MTB后的Mφ培养过程中逐渐增加。经奎纳克林或α-TFMK处理后，上述现象被阻断，表明AA以cPLA_2依赖的方式在吞噬体内转运到MTB。(3)IFN-γ激活的Mφ中cPLA_2和iNO_5mRNA表达增加，而sPLA_2mRNA表达无明显变化。(4)用抗cPLA_2抗体和芘标记的cPLA_2底物对感染的Mφ进行荧光显微镜观察，证实cPLA_2在Mφ的吞噬体中转位到分枝杆菌体内。(5)在小鼠腹腔Mφ中，在感染MTB后的早期培养阶段，可观察到MAP激酶依赖的cPLA_2丝氨酸残基磷酸化，至少在48 h内逐渐增加。另一方面，在未感染结核分枝杆菌的RAW Mφ中，cPLA_2的组成性磷酸化水平明显升高。这些结果表明cPLA_2在Mφ表达抗MTB和MAC的抗菌活性中起重要作用。

项目成果

Sato K, Tomioka H, Shimizu T, Gonda T, Ohta F, Sano C: "Type II alveolar cells roles in macrophage-mediated host innate resistance to pulumonary Mycobacterial infections by producing proinflammatory cytokines"The Journal of Infectious Diseases. 185. 1139-

Sato K、Tomioka H、Shimizu T、Gonda T、Ohta F、Sano C：“II 型肺泡细胞通过产生促炎细胞因子在巨噬细胞介导的宿主对肺部分枝杆菌感染的先天抵抗中发挥作用”《传染病杂志》。

冨岡 治明: "結核 (光山 正雄 編):分担執筆"医薬ジャーナル社. 322-333 (2001)

富冈晴明：“结核病（三山正夫编辑）：撰稿人”Iyaku Journalsha 322-333（2001）。

Sato K, Akaki T, Shimizu T, Sano C, Ogasawara K, Tomioka H: "Invasion and intracellular growth of Mycobacterium avium complex adapted to intramacrophage enviroment within macrophages and type II alveolar epitherial cells"Kekkaku. 76(2). 54-57 (2001)

Sato K、Akaki T、Shimizu T、Sano C、Ogasawara K、Tomioka H：“适应巨噬细胞和 II 型肺泡上皮细胞内巨噬细胞内环境的鸟分枝杆菌复合体的侵袭和细胞内生长”Kekkaku。

Sano C, Sano K, Sato K, Ogasawara K, Shimizu T, Tomioka H: "Effects of ATP on the in vivo and in vitro Antimicrobial activity of murine macrophages against Mycobacterium avium complex"Japanese J. Chemother. 50(12). 848-853 (2002)

Sano C、Sano K、Sato K、Ogasawara K、Shimizu T、Tomioka H：“ATP 对小鼠巨噬细胞体内和体外抗鸟分枝杆菌复合物抗菌活性的影响”Japan J. Chemother。

佐藤 勝昌, 赤木 竜也, 清水 利朗, 冨岡 治朗 他: "抗酸菌のマクロファージ(Mφ)と肺胞上皮細胞への感染性と細胞内増殖性:Mφ内順化菌を用いての検討"結核. 76・2. 54-57 (2001)

Katsumasa Sato、Tatsuya Akagi、Toshiro Shimizu、Jiro Tomioka 等：“抗酸细菌对巨噬细胞（Mφ）和肺泡上皮细胞的感染性和细胞内增殖：使用 Mφ 适应细菌的研究”结核病 76・2。 -57 (2001)